1-methyl-4-nitro-5-cyano-imidazole compositions for treating coccidiosis



United States Patent 1-METHYL-4-NITRO-5-CYANO-FMIDAZOLE COM- POSITIONS FOR TREATING COCCIDIOSIS Edward F. Rogers, Middletown, NJ., assignor to Merck & Co., Inc., Rahway, NJ in corporation of New Jersey No Drawing. Filed June 26, 1958, Ser. No. 744,671

3 Claims. (Cl. 167-531) the genus Eimeria, such as E. tenella, E. necatrix, E.,

acervulina, E. maxima, E. hagani and E. brunetti. E. tenella is the causative agent of a severe and often fatal infection of the caeca of chickens, which is manifested,

by severe and extensive hemorrhage, accumulation of blood in the caeca, and the passage of blood in the droppings. E. necatrix attacks the small 'mtestine of the chick.

causing what is known as intestinal coccidiosis. Related species of coccidia such as E. meleagridis and E. adenoidesare oaustive organismsof coccidiosis in turkeys. When left untreated, the severe forms of coccidiosis lead to poor weight gain, reduced feed efiiciency and high mortality in fowl. The elimination or control of this disease is, therefore, of paramount importance to the poultry raising industry.

I have now found that l-methyl-4-nitro-5-cyano imidazole is very active against protozoa which cause coccidiosis. As discussed more fully hereinbelow the development of coccidiosis is prevented when small amounts of this substance are fed to poultry. One object of the present invention, therefore, is to provide a new method for preventing and controlling coccidiosis. It is a further object to provide novel compositions which, when added to animal feedstuifs, are extremely effective in preventing coccidiosis. A still further object is provision of animal feedstutfs containing l-methyl-4-nitro-5-cyanoimidazole. Other objects will become apparent from the ensuing discussion of my invention.

1-methyl-4-nitro-S-oyano-imidazole has the structural formula N---o-N0 H02 1 ISO-0N In accordance with the present invention, it has been discovered that this material is a potent coccidiostat which may be successfully employed to prevent the development of coccidiosis when administered to poultry. My new coccidiostat is conveniently fed to poultry as a component of the'feed of the animals althoughit may also be given dissolved or suspended in the drinking water. According to a preferred aspect of the invention,-novel compositions for the treatment of coccidiosis are provided which comprise 1-methyl-4-nitro-5-cyano-imidazole intimately dispersed in or intimately admixed with an inert edible carrier or diluent. By an inert edible carrier or diluent is meant one that is nonreactive with respect to the imidazole, and that may be administered with safety to the animals to be treated. The carrier or diluent is 2,974,081 Patented Mar. 7, 1961 a 2 preferably one that is or may mal feed. g

The compositions which are a preferred feature of my invention are the so-called feed supplements in which. 1-methyl-4-nitro-5-cyano-imidazo1e is present in relatively large amounts and which are suitable for addition to the poultry feed either directly or after an intermediate dilution or blending step; Examples of carriers or diluents suitable for .such compositions are animal feed; ingredients such as distillers dried grains, corn meal,. citrus meal, fermentation residues, ground oyster shells, Attapul'gus clay, wheat shorts, molasses solubles, corrr cob meal, edible vegetable substances, toasted dehulled; soya flour, soybean mill feed, antibiotic mycelia, soya, grits, crushed limestone and the like. The imidazole. compound is intimately dispersed or admixed through-- be an ingredient of the ani-- out the solid inert carrier by methods such as grinding stirring, milling or tumbling. By selecting proper diluents and-by altering the ratio of carrier to active ingredient,. compositions of any desired concentration may be pre: pared. Formulations containing from about 1% to about 40% by'weight, and preferably fromab'out 225.% by weight, of 1-methyl-4-nitro-5-cyano-imidazole areparticularly suitablefor addition to poultry feedstuffs; those having from about 5-20% by weight of coccidiostat are very satisfactory. The active compound is usually dispersed or mixed uniformly in the diluent but in some instances may be sorbed on the carrier. Since it is conwenient for the feed-manufacturer to use about one pound of feed supplement for each ton of finished feed, the preferred concentration in the supplement is usually a function of the level of active ingredient desired in the finished feed.

Very low levels of 1-methyl-4-nitro-5-cyano-imidazole in an animal feed are sufiicient to afford the poultry good protection against coccidiosis. Preferably the compound:

is administered to chickens in an amount equal to about 0.0025 to about 0.025% by weight of the daily feed intake. Optimum results are obtained by feeding at a level of about 0.005% to 0.01% by weight of the finished feed.

For therapeutictreatment of an established coccidial infection, higher amounts of 1-methyl-4-nitro-5-cyanoimidazole, i.e. up to about 0.1% by weight of the feed consumed, may be employed. The most advantageous dosage level will, of course, vary somewhat with partic-' ular circumstances such as the type and severity of the coccidial infection to be treated.

For treating poultry, the feed supplement isiuniformly' dispersed in the animal feed by suitable mixing or-hlend-- ing procedures.

Usually the feed supplements are further diluted with materials such' as corn meal or soybean meal before being incorporated in' the animal feed; In this intermediate processing step the level of 1-methyl-4-nitro-5-cyanoimidazole in the carrier is brought down to about 0.1%

1.0% by weight. This dilution serves to facilitate uniform distribution of the coccidiostat in the finished, feed The finished feed is one that contains a source of fat,' protein, carbohydrate, minerals, vitamins and other nutritional factors.

In the above discussion of'my invention, emphasis has been placed on, solid compositions .wherein. the active ingredient is mixed with an ediblecarrier in a feedsup-- plement, in a so-called premix or'inthe final poultry" It will likewise be understood by those skilled in this art that special feed supplement formulations and finished animal feeds containing vitamins, antibiotics, growthpromoting agents and other nutritional substances may include the imidazole compound of this invention. A typical product of this type is the following:

Amount/lb. of

Distillers grains to 1 pound.

The following examples are given' for purposes of illustration and not by way of limitation:

Example 1 166 gramsof 1-methyl 4 nitro 5-ch1oro-imidaiole were dissolved in one liter of dry ethanol and to the resultingsolution wasadded 130 grams ofpotassium cyanide and 32 gramsof potassium'iodide. T he resultin'g mixture was refluxed with stirring for 151101115.

At the end of the reflux period, the mixturewascool'ed to room temperature and the precipitated saltsfiltered offi The clear filtrate wasconcentr'ated in 'vacuo to remove the alcohol. The residue of[1 methyl 4 nitro 5-cyanoimidazole thus obtained'wasrecrystallized from water to give sub'stantially pure material, melting point 141 C.

Example 2 Animal feed supplements having the fonowiag eoin positions at-e prepared brine-b nary i fl i f 'j' f nitro 5 cyano imidazole"- and" the particular edible" solid diluent or diluents:

A. 1-methyl-4-nitro-5-cyano imidaz'ole 7.5 Distillers dried grains 92.5

B. l-methyl-4-nitro-5-cyano-imidazole 5.0

Soybean mill fed 50.0 F1116 soya grits 45.0" c. 1-methyl 4-nitr'o-'-5-.cyano'-imidazole 1010;.

Molassessolubles 90.0

D. 1-methyl-4-nitro-S-cyano-imidazole 15.0 Corn distillers grains 55.0 Corn germ meal 30.0

E. l-methyl-4-nitro-5-cyano-imidazole 20.0 Wheat shorts 30.0 Distillers dried grains 50.0

F. 1-methyl-4-nitro-5-cyano-imidazole 25.0 Corn distillers dried grains 75.0

G. 1-methyl-4-nitro S-cyano-imidazole 40.0 Corn meal 60.0

H. 1-metliyl 4 nitro-5-cyano-imidazole 10.0 Nicarbazin 15.0 Corn distillers dried grains 75.0

These supplements are made by mechanical milling or mixing of the ingredients to insure uniform distribution of the active compound.

Example 3 Groups of three two-week old chick s were fed a mash diet containing 1-rnethyl-4-nitro-5-cyano imidazole uni formly dispersed in thefeed.. After having beetif on this ration for 24 hours, each bird was inoculated with 50,000 sporulated oocysts of E. tenella. Other groups of three two-week old chicks were fed a similar mash diet containingno imidazole. These were also infe'cted'after 24 hours and served as positive controls. Still other groups of three two-week old chicks were fed the mash diet free of l-methyl-4mitro 5-cyano-imidazole and were not infected with coccidiosis. v These served as normal controls. ,I I The experiment was completed on the seventh dayafter infection. During the seven-day period the infected birds were observed for clinical evidence'of coccidiosis. The surviving birds were weighed, sacrificed and examined for cecal coccidiosis. The following results were obtained:

Average Weight Percent (Gms.) Evidence Percent Morof Gooin Feed tallty cldlosls Initial Termlnal 0. 0025 17 165 moderate. 1 Methyl 4 nltro 5 0. 005 0 110 203 slight.

(ammo-imidazole. 0. 01 0 110 186 none. r 0. 02 0 113; 158. none. Positive Control 25 i 108 153 severe. N ormal' Control 0 206 nererhs cneu in thefile ofthis patent Beilst'ein': v61. 25 (II), 107. 

1. A COMPOSITION USEFUL IN THE PREVENTION AND TREATMENT OF COCCIDIOSIS WHICH COMPRISES 1-METHYL-4-NITRO-5-CYANOIMIDAZOLE IN INTIMATE DISPERSION IN AN ANIMAL FEEDSTUFF. 